title: Isoform characterisation, heterologous expression and functional analysis of two lectins from Vatairea macrocarpa.
abstract: VML is a lectin from Vatairea macrocarpa seeds that has various biological activities. Here, we describe three new lectin isoforms from V. macrocarpa identified through genomic DNA analysis. One of these isoforms has high similarity to VML, while another that has noteworthy differences. We have denoted the new isoforms as VML-2, VML-3 and VML-4. Recombinant VML (rVML) and VML-2 (rVML-2) were expressed in Escherichia coli and were anticipated to have similar biological activity compared to native VML. Recombinant lectins were produced using a synthetic gene strategy to improve the expression levels. We obtained two active recombinant lectin isoforms from V. macrocarpa, and there was no significant difference between their biological activities. The conservation between carbohydrate-binding sites of recombinant and native proteins was demonstrated by specific inhibition of hemagglutin activity by D-galactose and lactose. However, no inhibition was observed in the presence of glucose and mannose. Our data show that the recombinant lectins VML and VML-2 are active and capable of recognising D-galactose and lactose. Moreover, the absence of glycosylation does not interfere with their biological activity.
authors.affiliation: Laboratório de Moléculas Biologicamente Ativas (Biomol-Lab), Department of Biochemistry and Molecular Biology, Federal University of Ceará, Av. Humberto Monte s/n, Zip Code: 60455-970, P.O. Box 6043, Bloco 907, Lab. 1075, Campus do Pici, Fortaleza-CE, Brazil. bscavada@ufc.br.
authors.forename: João Garcia
authors.initials: JG
authors.lastname: Alves Filho
authors.forename: Antonia Sâmia Fernandes
authors.initials: AS
authors.lastname: do Nascimento
authors.forename: Ana Cláudia Silva
authors.initials: AC
authors.lastname: Gondim
authors.forename: Ronniery Hilario
authors.initials: RH
authors.lastname: Pereira
authors.forename: Rodrigo Maranguape Silva
authors.initials: RM
authors.lastname: da Cunha
authors.forename: Celso Shiniti
authors.initials: CS
authors.lastname: Nagano
authors.forename: Edson Holanda
authors.initials: EH
authors.lastname: Teixeira
authors.forename: Kyria Santiago
authors.initials: KS
authors.lastname: Nascimento
authors.forename: Benildo Sousa
authors.initials: BS
authors.lastname: Cavada
mesh_terms: D007887:Fabaceae / Q000737:chemistry* / Q000235:genetics; D005690:Galactose / Q000737:chemistry; D018506:Gene Expression Regulation, Plant; D018745:Genome, Plant; D008358:Mannose / Q000737:chemistry; D037121:Plant Lectins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D020033:Protein Isoforms / Q000737:chemistry / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D012639:Seeds / Q000737:chemistry
chemical_list: D037121:Plant Lectins; D020033:Protein Isoforms; D011994:Recombinant Proteins; D008358:Mannose; D005690:Galactose